Doering L C, Chang P L
Department of Biomedical Sciences (Anatomy) McMaster University, Hamilton, Ontario, Canada.
J Neurosci Res. 1991 Jul;29(3):292-8. doi: 10.1002/jnr.490290304.
Primary fibroblasts initiated from skin biopsies of Wistar rats were transfected with a plasmid that encodes the human growth hormone and the neomycin resistance genes. Cell clones selected for G418 resistance and expressing high levels of human growth hormone were propagated in vitro and subsequently transplanted into the cerebral cortex of adult allogeneic rats. Grafts were examined by immunocytochemistry at weekly intervals up to 2 months. Fibroblasts in the transplants survived and expressed growth hormone clearly up to 4 weeks, but showed reduced expression at 6 to 8 weeks. Significant levels of human growth hormone were also detected by radioimmunoassay in the serum of the host rats up to 1 month. The experiments demonstrate that primary fibroblasts can be genetically modified to deliver a new gene product into the central nervous system, and the gene product can pass the blood-brain barrier to enter the systemic circulation. This model illustrates the potential to introduce desired products into the brain through the genetic alteration of autologous primary fibroblasts.
从Wistar大鼠皮肤活检中分离出的原代成纤维细胞用编码人生长激素和新霉素抗性基因的质粒进行转染。选择对G418有抗性并表达高水平人生长激素的细胞克隆进行体外培养，随后移植到成年同种异体大鼠的大脑皮层中。在长达2个月的时间里，每周通过免疫细胞化学对移植物进行检查。移植的成纤维细胞存活下来，在4周内均清晰表达生长激素，但在6至8周时表达降低。通过放射免疫测定法在宿主大鼠血清中也检测到长达1个月的显著水平的人生长激素。这些实验表明，原代成纤维细胞可以通过基因改造将新的基因产物递送至中枢神经系统，并且该基因产物可以穿过血脑屏障进入体循环。该模型说明了通过自体原代成纤维细胞的基因改造将所需产物引入大脑的潜力。
